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Background Long-term survival in isolated marginal seas of the China coast during the late Pleistocene ice ages is widely believed to be an important historical factor contributing to population genetic structure in coastal marine species. Whether or not contemporary factors (e.g. long-distance dispersal via coastal currents) continue to shape diversity gradients in marine organisms with high dispersal capability remains poorly understood. Our aim was to explore how historical and contemporary factors influenced the genetic diversity and distribution of the brown alga Sargassum thunbergii, which can drift on surface water, leading to long-distance dispersal. Results We used 11 microsatellites and the plastid RuBisCo spacer to evaluate the genetic diversity of 22 Sargassum thunbergii populations sampled along the China coast. Population structure and differentiation was inferred based on genotype clustering and pairwise F ST and allele-frequency analyses. Integrated genetic analyses revealed two genetic clusters in S. thunbergii that dominated in the Yellow-Bohai Sea (YBS) and East China Sea (ECS) respectively. Higher levels of genetic diversity and variation were detected among populations in the YBS than in the ECS. Bayesian coalescent theory was used to estimate contemporary and historical gene flow. High levels of contemporary gene flow were detected from the YBS (north) to the ECS (south), whereas low levels of historical gene flow occurred between the two regions. Conclusions Our results suggest that the deep genetic divergence in S. thunbergii along the China coast may result from long-term geographic isolation during glacial periods. The dispersal of S. thunbergii driven by coastal currents may facilitate the admixture between southern and northern regimes. Our findings exemplify how both historical and contemporary forces are needed to understand phylogeographical patterns in coastal marine species with long-distance dispersal. Electronic supplementary material The online version of this article (10.1186/s12862-017-1089-6) contains supplementary material, which is available to authorized users.

William G North,1,2 Fuli Liu,1 Liz Z Lin,1 Ruiyang Tian,2 Bonnie Akerman1 1Department of Molecular and Systems Biology, Geisel School of Medicine at Dartmouth College, 2Woomera Therapeutics Inc, Lebanon, NH, USA Abstract: Pancreatic cancer, particularly adenocarcinoma of the pancreas, is a common disease with a poor prognosis. In this study, the importance of N-methyl-D-aspartate (NMDA) receptors for the growth and survival of pancreatic cancer was investigated. Immunohistochemistry performed with antibodies against GluN1 and GluN2B revealed that all invasive adenocarcinoma and neuroendocrine pancreatic tumors likely express these two NMDA receptor proteins. These proteins were found to be membrane components of pancreatic cancer cell lines, and both channel-blocker antagonist and GluN2B antagonist significantly reduced cell viability in vitro. Both types of antagonists caused an internalization of the receptors. Dizocilpine maleate (MK-801) and ifenprodil hemitartrate both significantly inhibited the growth of pancreatic tumor xenografts in nu/nu mice. These findings predict that, as for other solid tumors investigated by us, pancreatic cancer could be successfully treated, alone or in combination, with NMDA receptor antagonists or other receptor-inhibiting blocking agents. Keywords: pancreatic cancer, NMDA receptors, inhibitors, potential therapy

It has been previously reported that Rho‑kinase (ROCK) and poly ADP-ribose polymerase (PARP) serve critical roles in myocardial ischemia/reperfusion (I/R) injury. Studies have additionally demonstrated that the activation of ROCK and the expression of PARP are increased in I/R. However, the effect and mechanism of the two proteins remains to be fully elucidated in I/R. In addition, whether they can be influenced by each other is unclear. In the present study, it was demonstrated that ischemia followed by reperfusion resulted in a significant increase in ROCK and PARP. In addition, Y‑27632 (ROCK inhibitor) and 3‑aminobenzamide (3‑AB; PARP inhibitor) pretreatment rescued myocardial infarction size and cardiomyocyte apoptosis. The inhibitory role of Y‑27632 was observed to be superior to that of the 3‑AB group. In addition, Y‑27632 and 3‑AB diminished extracellular signal‑related kinase (ERK) phosphorylation and the production of tumor necrosis factor α and interleukin 6. Overall, the results of the present study suggested that the inhibition of ROCK leads to reduced myocardial infarction size and cardiomyocyte apoptosis via the PARP/ERK signaling pathway.

In this work, magnetic ferrite modified carbon nanotubes (MFe2O4/CNTs, M: Mn or Co) were synthesized and employed as adsorbents to remove emerging pollutant bezafibrate (BZF) from aqueous solution. The structural and surface properties of the prepared adsorbents were characterized, and the performances of the MFe2O4/CNTs were systematically investigated from adsorption kinetics and mechanistic points of view. Results showed that the fabricated MFe2O4/CNTs could integrate the advantages of CNTs and ferrite, thereby exhibiting an excellent adsorption performance and recyclability. The MnFe2O4/CNTs were more effective than CoFe2O4/CNTs for BZF adsorption. When the initial concentration of BZF was varied from 5 to 40 mg L−1, the maximum adsorption capacity of MnFe2O4/CNTs and CoFe2O4/CNTs increased from 14.8 to 33.4 mg g−1 and 8.1 to 27.8 mg g−1, respectively. Moreover, the Langmuir isotherm model and pseudo-second-order equation could elaborate well the adsorption of BZF on MFe2O4/CNTs. The thermodynamic analysis further revealed that the adsorptions were spontaneous processes. Based on measurements of surface and pore diffusion and the results of adsorption reactions, possible mechanisms were proposed to explain the adsorption process. This research indicates that the MFe2O4/CNTs are potentially applicable for the removal of BZF from aqueous solution.

Tissue homeostasis is maintained by differentiated progeny of residential stem cells. Both extrinsic signals and intrinsic factors play critical roles in the proliferation and differentiation of adult intestinal stem cells (ISCs). However, how extrinsic signals are transduced into ISCs still remains unclear. Here, we find that heparan sulfate (HS), a class of glycosaminoglycan (GAG) chains, negatively regulates progenitor proliferation and differentiation to maintain midgut homeostasis under physiological conditions. Interestingly, HS depletion in progenitors results in inactivation of Decapentaplegic (Dpp) signaling. Dpp signal inactivation in progenitors resembles HS-deficient intestines. Ectopic Dpp signaling completely rescued the defects caused by HS depletion. Taken together, these data demonstrate that HS is required for Dpp signaling to maintain midgut homeostasis. Our results provide insight into the regulatory mechanisms of how extrinsic signals are transduced into stem cells to regulate their proliferation and differentiation., Summary: Heparan sulfate chains in Drosophila adult intestinal stem cells negatively regulate stem cell proliferation and differentiation to control tissue homeostasis under physiological conditions.

Background Caulerpa lentillifera is one of the most important economic green macroalgae in the world. Increasing demand for consumption has led to the commercial cultivation of C. lentillifera in Japan and Vietnam in recent decades. Concomitant with the increase of C. lentillifera cultivation is a rise in disease. We hypothesise that epiphytes or other microorganisms outbreak at the C. lentillifera farm may be an important factor contributing to disease in C. lentillifera. The main aims are obtaining differences in the microbial community structure and diversity between healthy and diseased C. lentillifera and key epiphytes and other microorganisms affecting the differences through the results of high-throughput sequencing and bioinformatics analysis in the present study. Results A total of 14,050, 2479, and 941 operational taxonomic units (OTUs) were obtained from all samples using 16S rDNA, 18S rDNA, and internal transcribed spacer (ITS) high-throughput sequencing, respectively. 16S rDNA sequencing and 18S rDNA sequencing showed that microbial community diversity was higher in diseased C. lentillifera than in healthy C. lentillifera. Both PCoA results and UPGMA results indicated that the healthy and diseased algae samples have characteristically different microbial communities. The predominant prokaryotic phyla were Proteobacteria, Planctomycetes, Bacteroidetes, Cyanobacteria, Acidobacteria, Acidobacteria and Parcubacteria in all sequences. Chlorophyta was the most abundant eukaryotic phylum followed by Bacillariophyta based on 18S rDNA sequencing. Ascomycota was the dominant fungal phylum detected in healthy C. lentillifera based on ITS sequencing, whereas fungi was rare in diseased C. lentillifera, suggesting that Ascomycota was probably fungal endosymbiont in healthy C. lentillifera. There was a significantly higher abundance of Bacteroidetes, Cyanobacteria, Bacillariophyta, Ulvales and Tetraselmis in diseased C. lentillifera than in healthy C. lentillifera. Disease outbreaks significantly change carbohydrate metabolism, environmental information processing and genetic information processing of prokaryotic communities in C. lentillifera through predicted functional analyses using the Tax4Fun tool. Conclusions Bacteroidetes, Cyanobacteria, Bacillariophyta, Ulvales and Tetraselmis outbreak at the C. lentillifera farm sites was an important factor contributing to disease in C. lentillifera.

The current meta-analysis was designed to determine the relationship between executive function (EF) and empathy, as well as to identify any moderators. A search of Chinese and English databases yielded 18 studies and 67 effect sizes involving a total of 6006 participants. Results with the random effects model showed that EF was significantly positively correlated with empathy (r = .14, p < .001). Subgroup analysis showed that EF was more strongly related to cognitive empathy (r = .20, p < .001) than to affective empathy (r = .09, p = .03). Looking at the two dimensions of empathy, we further found that cognitive empathy is closely related to subcomponents of EF, including inhibitory control (r = .23, p < .001), working memory (r = .20, p < .001), and cognitive flexibility (r = .15, p = .036), while only affective empathy was closely related to inhibitory control (r = .12, p < .001). Results suggested that future research should consider that the relationship between empathy and EF varies depending on the division of specific subcomponents. This finding may help in explaining possible mechanisms of how EF affects empathy.

Carbapenem-resistant Klebsiella pneumoniae (CRKP), one of the major nosocomial pathogens, is increasingly becoming a serious threat to global public health. There is an urgent need to develop effective therapeutic and preventive approaches to combat the pathogen. Here, we identified and characterized a novel capsule depolymerase (K64-ORF41) derived from Klebsiella phage SH-KP152410, which showed specific activities for K. pneumoniae K64-serotype. We showed that this depolymerase could be used in the identification of K64 serotypes based on the capsular typing, and the results agreed well with those from the conventional serotyping method using antisera. From this study, we also identified K64 mutant strains, which showed typing discrepancy between wzi-sequencing based genotyping and depolymerase-based or antiserum-based typing methods. Further investigation indicated that the mutant strain has an insertion sequence (IS) in wcaJ, which led to the alteration of the capsular serotype structure. We further demonstrated that K64-ORF41 depolymerase could sensitize the bacteria to serum or neutrophil killing by degrading the capsular polysaccharide. In summary, the identified K64 depolymerase proves to be an accurate and reliable tool for capsular typing, which will facilitate the preventive intervention such as vaccine development. In addition, the polymerase may represent a potential and promising therapeutic biologics against CRKP-K64 infections.

A novel wallboard with double layers shape-stabilized phase change materials (SSPCMs) for year round energy management was proposed. Its energy saving potential in five typical climate regions of China was studied. One office room integrated with double layers SSPCMs wallboard was used for simulation platform. The other normal office room with the same envelope and just without double layers SSPCMs wallboard was used as reference room. Studies were conducted to investigate the energy saving potential of double layers SSPCMs wallboard in five cities representing five typical climate regions of China, including severe cold region, cold region, hot summer and cold winter region, hot summer and warm winter region, mild region. The optimal melting temperature and corresponding thickness of SSPCMs wallboard in five representative cities were given when the energy saving potential reached maximal value. It concluded that the optimal melting temperature of internal SSPCM layer was effected by indoor air temperature and the optimal melting temperature of external SSPCM layer was effected by outdoor air temperature. The characteristic of optimal values in the five representative cities were given for reference in this paper.

A novel sandwich-type wallboard was that of a three-layer panel with external and internal layer consisting of shape-stabilized phase change materials (SSPCMs) wallboards and middle layer consisting of concrete. The external layer was active in hot seasons and the internal layer was active in cold seasons. One office room integrated with double SSPCMs wallboards was used for simulation platform. Another office room with the same envelope and just without double SSPCMs wallboards was used as reference room for comparison. Split variable and constant frequency air-conditioner were used in these two different rooms. Studies were conducted to investigate the effects of double SSPCMs and variable/constant frequency air-conditioner on energy performance in typical climate area with hot summer and cold winter (Wuhan city, China). Test results showed that building operation energy consumption in SSPCM room was 6.4% lower than that in reference room under variable frequency air-conditioner in summer. The building operation energy consumption in SSPCM room was 17.8% lower than that in reference room under variable frequency air-conditioner in winter. It concluded that the effect of variable frequency air-conditioner used in SSPCM room was better than that in reference room both in summer and winter. But the energy saving effect was more obvious in winter than that in summer.

Pyropia dentata is a promising species for aquaculture in South China. To develop an efficient protocol for seedling culture, the growth, development, and physiological changes of P. dentata conchocelis were investigated under different temperatures, irradiances, photoperiods, and salinity. After 25 days, conchosporangia formation at 28 °C was 5–7-fold of that at 25 and 31 °C. The percentage of conchosporangia increased as day length increased from 8 to 12 h and then decreased sharply under day length ≥14 h, with no conchosporangia observed under 24L:0D. By contrast, long day length (≥14 h) and lower temperature (≤25 °C) favored the vegetative growth of conchocelis. Conchosporangia formation peaked at 28 °C, 12L:12D, and 40–60 μmol photons m−2 s−1. The highest F v /F m , P n , and P n /R d also occurred at 28 °C and/or 12L:12D, which might provide energy for conchosporangia formation. Total soluble proteins (TSPs) content decreased while relative growth rate increased as irradiance and day length increased. Increasing irradiance and day length, and at temperatures of 31 °C caused a reduction in phycoerythrin and chlorophyll a content. However, allophycocyanin content increased at 31 °C and the content of phycocyanin, carotenoids, and TSPs remained steady from 25 to 31 °C. Growth, development, P n , content of TSPs and pigments were similar at 25–40 ppt. F v /F m decreased and R d increased significantly at salinities ≥35 ppt. These findings provide informative data on the mechanisms of Pyropia conchocelis development, and help establish an efficient seedling culture protocol for this P. dentata strain.

Organic UV filters (OUV-Fs) are increasingly used for protection against UV irradiation. The widespread occurrence of OUV-Fs residues in aquatic systems has been reported, but little is known regarding their distribution and potential impact to the surface water in China. This study reports the occurrence, distribution and risk assessment of eight widely used OUV-Fs in the surface water of Nanjing. The results indicated butyl-methoxydibenzoylmethane, octyl-dimethyl-PABA and benzophenone-types (BP3, BP4 and BP1) were the most frequently detected compounds at concentrations of 3.63-104 ng/L. In general, the concentrations of OUV-Fs were decreased along the rivers; however, due to a substantial pollution load from the tributaries, higher concentrations of OUV-Fs were observed near the tributary inlet, compared to the other study areas. The risk assessment based on the calculated risk quotients (RQs) demonstrated that all OUV-Fs posed at least low risks to certain sensitive aquatic organisms, and BP3 posed high risk with RQ values of 1.64. It should be noted that the exclusion of adsorbed OUV-Fs might have contributed to an underestimation of the risk, therefore, it's necessary to assess both adsorbed and dissolved OUV-Fs in further studies.

This study aimed to investigate the protective effect of GanodericacidA (GA) on myocardial ischemia-reperfusion (MIR) injury. The myocardial injury model in rats was established by ligating left anterior descending coronary artery. We measured cardiac hemodynamic, antioxidant enzyme activity, and various biochemical indexes of myocardial tissue, and evaluated myocardial infarction and damage. Further, the expression of JAK2/STAT3/NF-κB signaling pathway-related proteins in myocardial tissue was measured by western blot. The results showed that the myocardial infarction extention was obviously reduced upon GA treatment. Compared with the control group, ischemia-reperfusion rats showed significant increase in lactate dehydrogenase (LDH) and creatine Kinase (CK), which were significantly decreased in GA group. Besides, GA pretreatment effectively decreased the levels of inflammatory cytokines in serum. The phosphorylation of Janus Kinase 2 (JAK2), signal transducer and activator of transcription (STAT3)and Nuclear factor-κB (NF-κB) in reperfusion group were significantly higher than that in control group, which were reversed upon GA treatment. In conclusion, GA may reduce myocardial injury by regulating JAK2/STAT3/NF-κB pathway.

To investigate the distribution characteristics of typical toxic elements in different mineralogical phases of fly ash is of significance when fly ash is comprehensively utilized. In this study, lignite fly ash can be preliminarily separated into three mineralogical phases: unburned lignite, iron microbeads and aluminate-silicate microbeads by two methods namely screening and dry magnetic separation. Then, the aluminate-silicate microbeads were subjected to two-step leaching. The first step was to investigate whether toxic elements migrated easily in the environment by column leaching test. In the second step, the aluminate-silicate microbeads were stripped from the surface of the particles to the internal by the acid-base combined leaching method, then the structural characteristics of the product and the trend of toxic elements content were explored. The results showed that there were few toxic elements in unburned lignite and the toxic elements Cr, Ni, Mo and Cd had a relatively high proportion in the iron microbeads. Column leaching results showed that the toxic elements V, Cr, Mn, Co, Cu, Hg and Pb had higher leaching rates, which proved that these elements were significantly enriched on the surface of the particles and easily migrated in the environment. Cr, Mo, Cd and W were highly enriched in the quartz-mullite mixture. Mn, Co, Ni, Cu, Zn and As were highly enriched in the amorphous component. The toxic elements exhibited different leaching rules during the acid-base combined leaching process revealing the complex embedded relationship with constant elements.

The structure of a new wall system was that of a three-layer sandwich-type panel with external/internal layer consisting of shape-stabilized phase change material (SSPCM) wallboard and middle layer consisting of conventional concrete. The external layer was active in hot seasons and the internal layer was active in cold seasons. An office room integrated with the novel double SSPCMs wallboard with a splitting air-conditioner was used for modeling and simulation test platform. Studies were conducted to investigate the effects of the novel double SSPCMs and different parameters on the energy and thermal performance in typical climate area with hot summer and cold winter (Wuhan city, China). Test results showed that the recommended thicknesses of external and internal SSPCM wallboard were 30–60 mm. With the recommended thickness, the annual energy savings for cooling were 3.4–3.9%. The peak load reductions for cooling were 3.1–3.8%. The annual energy savings for heating were 14.8–18.8%. The peak load reductions for heating were 8.6–11.3%. The external layer was effective on reducing annual energy demand and peak load of cooling in hot seasons. The internal layer was effective on reducing the indoor temperature fluctuation in cold seasons.

William G North, Fuli Liu, Ruiyang Tian, Hamza Abbasi, Bonnie Akerman Department of Physiology and Neurobiology, Geisel School of Medicine at Dartmouth College, Lebanon, NH, USA Abstract: We have earlier demonstrated that breast cancer and small-cell lung cancer express functional NMDA receptors that can be targeted to promote cancer cell death. Human ovarian cancer tissues and human ovarian cancer cell lines (SKOV3, A2008, and A2780) have now been shown to also express NMDA-receptor subunit 1 (GluN1) and subunit 2B (GluN2B). Seventeen ovarian cancers in two arrays were screened by immunohistochemistry using polyclonal antibodies that recognize an extracellular moiety on GluN1 and on GluN2B. These specimens comprised malignant tissue with pathology diagnoses of serous papillary cystadenocarcinoma, endometrioid adenocarcinoma, and clear-cell carcinoma. Additionally, archival tissues defined as ovarian adenocarcinoma from ten patients treated at this institute were also evaluated. All of the cancerous tissues demonstrated positive staining patterns with the NMDA-receptor antibodies, while no staining was found for tumor-adjacent normal tissues or sections of normal ovarian tissue. Human ovarian adenocarcinoma cell lines (A2008, A2780, SKOV3) were demonstrated to express GluN1 by Western blotting, but displayed different levels of expression. Through immunocytochemistry utilizing GluN1 antibodies and imaging using a confocal microscope, we were able to demonstrate that GluN1 protein is expressed on the surface of these cells. In addition to these findings, GluN2B protein was demonstrated to be expressed using polyclonal antibodies against this protein. Treatment of all ovarian cell lines with antibodies against GluN1 was found to result in decreased cell viability (P

Pyropia germplasm qualified for drift system culture is highly applicable in China. The current farmed species are tolerant to abiotic stresses, but vulnerable without air drying. Pyropia katadae var. hemiphylla inhabits mid-intertidal marshes and is a candidate for drift-system farming. Here, the performance of the gametophytes under varying environmental conditions was investigated. The thalli endured a short period (

As a temperate-cold species, Saccharina japonica often suffers heat stress when it is transplanted to temperate and subtropical zones. Study the heat stress response and resistance mechanism of Saccharina is of great significance for understanding the acclimation to heat stress under domestication as well as for breeding new cultivars with heat stress resistance. In this study, we identified a set of heat stress-responsive miRNAs and analysed their regulation during the heat stress response. CO (control) and heat stress (HS) sRNA libraries were constructed and sequenced. Forty-nine known miRNAs and 75 novel miRNAs were identified, of which seven known and 25 novel miRNAs were expressed differentially under heat stress. Quantitative PCR of six selected miRNAs confirmed that these loci were responsive to heat stress. Thirty-nine and 712 genes were predicted to be targeted by the seven known miRNAs and 25 novel miRNAs, respectively. Gene function and pathway analyses showed that these genes probably play important roles in S. japonica heat stress tolerance. The miRNAs identified represent the first set of heat-responsive miRNAs identified from S. japonica, and their identification can help elucidate the heat stress response and resistance mechanisms in S. japonica.

Pharmaceutically active compounds (PhACs) have been universally detected in rivers, lakes and coastal waters that are affected by effluents from sewage treatment plants (STPs). In this study, the biological effects and bioaccumulation of PhACs were assessed in crucian carp (Carassius auratus) caged in an effluent-receiving river for 21 days. Compared with control fish in the laboratory and at a reference site, a significant reduction in hepatosomatic index (HSI) and increase in the biotransformation enzymes ethoxyresorufin O-deethylase (EROD) and glutathione S-transferase (GST) activities were observed in the fish that was caged downstream from the STP outfall. In general, the total concentrations of PhACs detected in fish tissues were in the order as follows: liver > brain > gill > muscle > bile. The bioaccumulation factors (BAFs) for PhACs were between 195 and 2782 in the major storage tissue liver. The corresponding results for both risk quotient (RQ) and enhanced integrated biomarker response (EIBR) based on laboratory and field studies, respectively, indicated that environmental risk for adverse effects to aquatic organisms were clearly higher at the downstream of the STP outfall than at the upstream.

Saccharina japonica is a commercially and ecologically important seaweed and is an excellent system for understanding the effects of domestication on marine crops. In this study, we used 19 selected simple sequence repeat (SSR) markers to investigate the influence of domestication on the genetic diversity and structure of S. japonica populations. Wild kelp populations exhibited higher genetic diversity than cultivated populations based on total NA, HE, HO, NP and AR. Discriminant analysis of principal components (DAPC), a neighbour-joining (NJ) tree and STRUCTURE analyses indicated that S. japonica populations could be divided into two groups (a cultivated/introduced group and a wild indigenous group) with significant genetic differentiation (P S. japonica has not resulted in any serious genetic disturbance of wild introduced S. japonica populations. An understanding of the genetic diversity and genetic structure of domesticated S. japonica will be necessary for further genetic improvement and effective use of germplasm.

International audience; The sporophyte generation of the brown alga Ectocarpus sp. exhibits an unusual pattern of development compared to the majority of brown algae. The first cell division is symmetrical and the apical/basal axis is established late in development. In the IMMEDIATE UPRIGHT mutant the initial cell undergoes an asymmetric division to immediately establish the apical/basal axis. We provide evidence which suggests that this phenotype corresponds to the ancestral state of the sporophyte. The IMM gene encodes a protein of unknown function, containing a repeated motif also found in the EsV-1-7 gene of the Ectocarpus virus. Brown algae possess large families of EsV-1-7 domain genes but these genes are rare in other stramenopiles suggesting that the expansion of this family may have been linked with the emergence of multicellular complexity. EsV-1-7 domain genes have a patchy distribution across eukaryotic supergroups and occur in several viral genomes, suggesting possible horizontal transfer during eukaryote evolution.

Enediyne natural products are potent antibiotics structurally characterized by an enediyne core containing two acetylenic groups conjugated to a double bond in a 9- or 10-membered carbocycle. The biosynthetic gene clusters for enediynes encode a novel iterative type I polyketide synthase (PKSE), which is generally believed to initiate the biosynthetic process of enediyne cores. This review article will cover research efforts made since its discovery to elucidate the role of the PKSE in enediyne core biosynthesis. Topics covered include the unique domain architecture, identification, and characterization of turnover products, and interaction with partner thioesterase protein. © 2014 IUBMB Life, 66(9):587–595, 2014

New variety breeding is very significant for Saccharina japonica cultivation industry. In this paper, we reported the breeding process, evaluation of economic traits, and commercial cultivation of a new Saccharina variety “Huangguan No. 1” (variety approval number: GS-01-006-2011). The sporophytes with best performance were screened out from varieties cultivated throughout Fujian Province, China, and mixed together as the parent population. The “Huangguan No. 1” variety was bred successfully from 5 years of targeted selection of objective traits and continuous progeny selfing/inbreeding. This new variety has a much longer, wider, and thicker blade; faster growth rate; heavier individual weight; and stronger resistance to high temperatures compared to the control variety (p

Sargassum thunbergii is an economically important brown alga that is used as a perferred food for sea cucumber in China. However, reports on the reproductive biology of S. thunbergii are limited. This study observed the characteristics of mature receptacles. The effects of different temperatures, light intensities, and photoperiods on the egg release of mature S. thunbergii receptacles were investigated. A liquid-phase oxygen electrode system was used to obtain light saturation and light compensation points of egg and young thalli of S. thunbergii. Results showed that temperature was the key factor for the egg release of mature receptacles. The conditions most conducive to egg release were 20&degC to 23&degC temperature and 50 μmol photons m-2&middots-1 to 200 μmol photons m-2&middots-1 light intensity. In addition, the light saturation and compensation points of unfertilized eggs or young thalli at 2 d or 3 d postfertilization ranged from 90 μmol photons m-2&middots-1 to 120 μmol photons m-2&middots-1 and 14 μmol photons m-2&middots-1 to 22 μmol photons m-2&middots-1, respectively. The combined results provide a reference for the sporeling culture of S. thunbergii.

Saccharina japonica is a common macroalga in sublittoral communities of cold seawater environments, and consequently may have highly efficient ribulose-1, 5-bisphosphate carboxylase/oxygenase (Rubisco) activity for carbon assimilation. In our study, we cloned the full-length Rubisco gene from S. japonica (SJ-rbc). It contained an open reading frame for a large subunit gene (SJ — rbcL) of 1 467 bp, a small subunit gene (SJ-rbcS) of 420 bp, and a SJ-rbcL/S intergenic spacer of 269 bp. The deduced peptides of SJ-rbcL and SJ-rbcS were 488 and 139 amino acids with theoretical molecular weights and isoelectric points of 53.97 kDa, 5.81 and 15.84 kDa, 4.71, respectively. After induction with 1 mmol/L isopropyl-β-D-thiogalactopyranoside for 5 h and purification by Ni2+ affinity chromatography, electrophoresis and western blot detection demonstrated successful expression of the 55 kDa SJ-rbcL protein. Real-time quantitative PCR showed that the mRNA levels of SJ-rbcL in gametophytes increased when transferred into normal growth conditions and exhibited diurnal variations: increased expression during the day but suppressed expression at night. This observation implied that Rubisco played a role in normal gametophytic growth and development. In juvenile sporophytes, mRNA levels of SJ-rbcL, carbonic anhydrase, Calvin-Benson-Bassham cycle-related enzyme, and chloroplast light-harvesting protein were remarkably increased under continuous light irradiance. Similarly, expression of these genes was up-regulated under blue light irradiance at 350 μmol/(m2·s). Our results indicate that long-term white light and short-term blue light irradiance enhances juvenile sporophytic growth by synergistic effects of various photosynthetic elements.

Saccharina japonica is a cold-temperate species; it often suffers heat stress during cultivation in temperate and subtropical zone in China. In this study we investigated the response of this alga to heat stress on transcriptomic level. A total of 947 genes (1.32 %) were identified as different expression genes (DEGs), out of which 548 and 399 genes were respectively up- and down-regulated by the heat stress. Among the 138 enriched gene ontology (GO) terms, “cell part”, “binding” and “cellular process” ranked as the first three GO terms that contained the most DEGs. A total of 47 pathways involved 119 DEGs were enriched. Over half of these DEGs were involved in “protein processing in endoplasmic reticulum”, “metabolic pathway” and “biosynthesis of secondary metabolites”. Manual classification of DEGs indicated that 155 DEGs were divided into nine groups, including heat shock protein, antioxidant system, protein synthesis and degradation, and so on. The results indicated that the heat stress triggered complex response of S. japonica; in turn, S. japonica adjusted its physiological and metabolic processes to adapt to and survive the heat stress. The identified DEGs will shed light on the heat tolerance mechanism of S. japonica and benefit future breeding of variety with heat tolerance.

Sargassum thunbergii has been studied extensively because of its particular ecology and economic value. Unfortunately, further progress has been hampered by lack of available genomic information. In this study, we sequenced and de novo assembled the transcriptome of S. thunbergii. A total of 53,490,957 clean reads were produced and were assembled into 46,269 contigs with the length from 351 nt to 21,538 nt and an average of 1,130 nt. Open reading frame (ORF) prediction in the assembled sequences showed that 30,139 (65.1 %) contigs had ORFs, and BLAST analysis indicated that 28,667 (61.96 %) sequences were annotated with putative functions. A total of 15,166 (32.8 %) putative genes were assigned at least one gene ontology term and 4,263 (10.0 %) putative proteins were classified into 25 categories by aligning against COG/KOG database. Function annotation with KEGG database assigned 11,798 (25.5 %) putative genes to 299 pathways. Out of the six enzymes that may be involved in alginate biosynthesis pathway, four have been identified from the transcriptome. Two pathways of guanosine diphosphate-mannose synthesis were identified in this alga, and the genes encoding the three putative enzymes responsible for mannitol cycle were discovered. Moreover, about 209 putative genes encoding the different kinds of heat shock proteins and a total of 119 putative genes encoding kinds of reactive oxygen species scavengers were also identified in the transcriptome, indicating that S. thunbergii may have strong resistance or tolerance to various stresses. In short, to our knowledge, this is the first data set for S. thunbergii transcriptome and will be a valuable resource for further studies on this alga.

Sargassum thunbergii is of great economic and ecological value to sea cucumber cultures and seaweed beds. However, studies on photosynthesis and respiration of S. thunbergii are limited. In this study, a liquid-phase oxygen electrode system and a chlorophyll fluorescence spectrometer were used to determine the photosynthetic characteristics of S. thunbergii seedlings at various light intensities, temperatures, and salinities. The light-saturated net photosynthetic rates, light saturation points, and respiratory rates of germlings were investigated. Results showed that the increase in rate of light saturation point was slow in the first 4 d, rapidly increased from 5 d to 9 d, and then gently increased at the final few days of the 15 d indoor culture period. The photosynthetic rate or respiratory rate of the seedlings rapidly and significantly decreased when the temperature was &degC or >28&degC. Short-term high- or low-salinity shock had significant effect on the photosynthetic and respiratory rates of the seedlings, specifically at 10 and 50 psu. However, the photosynthetic and respiratory rates recovered to the normal levels after 24 h of recovery period, which demonstrated a powerful ion-transport system of the seedlings. These results provided reference for the artificial breeding of S. thunbergii.

Radar signal sorting is a key part of radar reconnaissance. A blind separation algorithm based on detection of time frequency is proposed in this paper and the problems of underdetermined radar signal sorting can be solved effectively. Firstly, the method is that single source point of each radar source signal was detected. Then; the mixing vector in the corresponding single source point set was estimated by Singular Value Decomposition (SVD). Finally, the mixing matrix simultaneously were estimated by the cluster validation technique based on k-means clustering algorithm, and the radar signals can be got by the mixing matrix and the observed signals. Each time domain waveform of radar source can be sorted based on this method, the time frequency graphs of radar signal can be got and the whole radar signal sorting process is accomplished.

To address the problems of “bottleneck” of expert system (ES) in acquiring knowledge as well as “match conflict” and “combinational explosion” in expert system inference, base on fundamental principles of Maintainability appraisal of electronic recycling equipment in a type of missile, the diagnostic method is proposed for locating the fault of module unit or key component in various PCB Maintainability appraisal models by embedding SVM into ES inference engine, by which the design incorporates construction of ES inference engine diagnosing the faults with electronic recycling equipment in a type of missile based on SVM, together with the method for realization of SVM inference engine. The experiments prove that this method effectively improves ES fault diagnosis rate and identification ratio.

The history of English test reform on secondary level in China is a reflection of Chinese development in recent 30 years. The stress on English language helps move on Chinese reform-and-open process. On the new historical step, there are some criticisms of English language and English language education in China. Particularly some people hold that it is necessary to reform the English tests, to re-understand the significance of English, and to lower down its weight in education. Meanwhile some others still emphasize the importance of English language and English language education in China, refusing to weaken the status of English in school education. The two trends of thinking could be explained by the cultural politics of English education in mainland China. Though English helps in Chinese reform-and-open history, it produces difficulty in education because of its role as a foreign language, and it would bring about some possible problem in the field of ideology.

To establish a molecular-marker-assisted system of breeding and genetic study for Laminaria japonica Aresch., amplified fragment length polymorphism (AFLP) was used to construct a genetic linkage map of L. japonica featuring 230 progeny of F2 cross population. Eighteen primer combinations produced 370 polymorphic loci and 215 polymorphic loci segregated in a 3:1 Mendelian segregation ratio (P ≤ 0.05). Of the 215 segregated loci, 142 were ordered into 27 linkage groups. The length of the linkage groups ranged from 6.7 to 90.3 centimorgans (cM) with an average length of 49.6 cM, and the total length was 1,085.8 cM, which covered 68.4% of the estimated 1,586.9 cM genome. The number of mapped markers on each linkage group ranged from 2 to 12, averaging 5.3 markers per group. The average density of the markers was 1 per 9.4 cM. Based on the marker density and the resolution of the map, the constructed linkage map can satisfy the need for quantitative trait locus (QTL) location and molecular-marker-assisted breeding for Laminaria.

Using transcriptome data to mine microsatellite and develop markers has growingly become prevalent. However, characterizing the possible function of microsatellite is relatively rare. In this study, we explored microsatellites in the transcriptome of the brown alga Sargassum thunbergii and characterized the frequencies, distribution, function and evolution and developed primers to validate these microsatellites. Our results showed that Tri-nucleotide is the most abundant, followed by di- and mono-nucleotide. The length of microsatellite was significantly affected by the repeat motif size. The density of microsatellite in the CDS region is significantly lower than that in the UTR region. The annotation of the transcripts containing microsatellite showed that 573 transcripts have GO terms and can be categorized into 42 groups. Pathways enrichment showed that microsatellites were significantly overrepresented in the genes involved in pathways such as Ubiquitin mediated proteolysis, RNA degradation, Spliceosome, etc. Primers flanking 961 microsatellite loci were designed and among the 30 pairs of primer selected randomly for availability test, 23 were proved to be efficient. These findings provided new insight into the function and evolution of microsatellite in transcriptome and the identified microsatellite loci within the annotated gene will be useful for developing functional markers in S. thunbergii.

In the present study, we selected the sporophytes of eight representative cultivars and four wild populations as specimens to assess their genetic diversity level and investigate their population genetic structure using the simple sequence repeat (SSR) markers. The genetic diversity analysis showed that the average genetic diversity of the wild populations from Russia Far East was higher than that of the wild populations from Northern Japan, which was in turn higher than that of the cultivars from China. The wild populations harboring higher genetic diversity level than the cultivars implied that S. japonica domestication might be accompanied by genetic diversity decrease. The four wild populations and the eight cultivars were taken as two groups respectively. Analysis of molecular variance (AMOVA) results indicated that there was significant genetic differentiation between the two groups (Phi(CT)=0.212, P

In the present study, we investigated whether omentin affected the expression of intracellular adhesion molecule-1 (ICAM-1) and vascular cell adhesion molecule-1 (VCAM-1) in tumor necrosis factor-α (TNF-α) induced human umbilical vein endothelial cells (HUVECs). Our data showed that omentin decreased TNF-α-induced expression of ICAM-1 and VCAM-1 in HUVECs. In addition, omentin inhibited TNF-α-induced adhesion of THP-1 cells to HUVECs. Further, we found that omentin inhibited TNF-α-activated signal pathway of nuclear factor-κB (NF-κB) by preventing NF-κB inhibitory protein (IκBα) degradation and NF-κB/DNA binding activity. Omentin pretreatment significantly inhibited TNF-α-induced ERK activity and ERK phosphorylation in HUVECs. Pretreatment with PD98059 suppressed TNF-α-induced NF-κB activity. Omentin, NF-kB inhibitor (BAY11-7082) and ERK inhibitor (PD98059) reduced the up-regulation of ICAM-1 and VCAM-1 induced by TNF-α. These results suggest that omentin may inhibit TNF-α-induced expression of adhesion molecules in endothelial cells via blocking ERK/NF-κB pathway.

The public availability of numerous expressed sequence tag (EST) enables EST-based SSR (simple sequence repeat) markers to be widely used for genetics and breeding studies. In the present study, EST-SSR markers were developed from ESTs of and were transferred to the non-congeneric species . Among the 2,668 non-redundant ESTs, 83 (3.1%) ESTs containing SSR were identified totally, with an average of one SSR per 13.6 kb. Analysis of SSR motifs revealed that the trinucleotide and tetranucleotide were major motifs, accounted for 44.58% and 16.87%, respectively. Based on the 83 ESTs containing SSR, we designed 45 pairs of primers in the flanking regions of the SSR, of which 13 pairs showed polymorphism in a wild population, and the mean alleles per locus was 3.6 (ranging from 2 to 6). The observed (Ho) and expected (He) heterozygosities of these EST-SSRs were 0.234-0.632 and 0.260-0.635, respectively. All loci were in Hardy-Weinberg equilibrium in the wild population and no linkage disequilibrium was detected among loci. The obtained EST-SSR markers can facilitate and promote related research such as ecological investigation, genetic diversity assessment and breeding practice of as well.

Chemical modulators of autophagy provide useful pharmacological tools for examination of autophagic processes, and also may lead to new therapeutic agents for diseases in which control of cellular sequestration and degradation capacity are beneficial. We have identified that timosaponin A-III (TAIII), a medicinal saponin reported to exhibit anticancer properties and improve brain function, is a pronounced activator of autophagy. In this work, the salient features and functional role of TAIII-induced autophagy were investigated. In TAIII-treated cells, autophagic flux with increased formation of autophagosomes and conversion into autolysosomes is induced in association with inhibition of mammalian target of rapamycin activity and elevation of cytosolic free calcium. The TAIII-induced autophagy is distinct from conventional induction by rapamycin, exhibiting large autophagic vacuoles that appear to contain significant contents of endosomal membranes and multivesicular bodies. Furthermore, TAIII stimulates biosynthesis of cholesterol, which is incorporated to the autophagic vacuole membranes. The TAIII-induced autophagic vacuoles capture ubiquitinated proteins, and in proteasome-inhibited cells TAIII promotes autophagy of aggregation-prone ubiquitinated proteins. Our studies demonstrate that TAIII induced a distinct form of autophagy, and one of its pharmacological actions is likely to enhance the cellular quality control capacity via autophagic clearance of otherwise accumulated ubiquitinated protein aggregates., link_to_subscribed_fulltext

To construct a molecular-marker-assisted selection (MAS) system, research was done on identifying molecular markers linking to longer frond length, a crucial selection index in the breeding of the commercially important seaweed Saccharina japonica. An F2-segregant population of 92 individuals was obtained by crossing two prominent S. japonica strains. Genomic DNA from ten individuals with the longest frond and ten individuals with the shortest frond in the F2-segregant population were mixed to create two DNA pools for screening polymorphic markers. In bulked-segregant analysis (BSA), out of 100 random amplified polymorphic DNA (RAPD) primers only two produced three polymorphic RAPD markers between the two DNA pools. In conversion of the three RAPD markers into sequence-characterized amplified region (SCAR) markers, only one was successfully converted into a SCAR marker FL-569 linking to the trait of longer frond. Test of the marker FL-569 showed that 80% of the individuals with longest fronds in a wild population and 87.5% of individuals with the longest fronds in an inbred line “Zhongke No. 2” could be detected by FL-569. Additionally, genetic linkage analysis showed that the SCAR marker could be integrated into the reported genetic map and QTL mapping showed that FL-569 linking to qL1-1. The obtained marker FL-569 will be beneficial to MAS in S. japonica breeding.

Unraveling the cryptic genetic diversity and selective breeding network in various Porphyra strains is of significance for conservation and utilization of economically important nori crops, for both current and future needs. Here, we used nuclear ribosomal spacer (ITS1) region to investigate the genetic variation and intra-specific relatedness of 59 Porphyra yezoensis Ueda specimens worldwide using phylogenetic and parsimony genealogical approaches. 23 nrDNA ITS1 genotypes were revealed and clustering analysis grouped them into two distinct clades. High genetic diversity was detected in wild P. yezoensis strains from Miyagi and Hokkaido Prefectures in Japan, while the cultivated strains from China and South Korea exhibited relatively higher genetic diversity likewise, which provided crucial genetic insights for future commercial breeding of P. yezoensis on a global scale. In addition, phylogenetic study has revealed the genetic relationship of strains with unknown parentage to those with known parentage, and also ITS1 sequence pattern could be correlated with the geographic origin of P. yezoensis specimens. All these pedigree information generated from this research can be used to select parents for inter-specific or intra-specific selective breeding and cross breeding to maximize the preservation of stock resource and sustainable development of nori industry.

Heat shock protein 70 (Hsp70) is one of the important members of heat shock protein (Hsp) families and plays essential roles in folding nascent protein, translocation, refolding denatured protein, protein degradation, adverse stress resistance, and so on. In this study, homologous cloning coupled with the rapid amplification of cDNA ends was used to clone full-length cytosolic heat shock protein 70 of Enteromorpha prolifera (designed as EPHsp70). Bioinformatics was used to analyze structural feature, homologous relationship, and phylogenetic position of EPHsp70. The full length of EPHsp70 cDNA was 2,265 bp, with a 5' untranslated region of 65 bp, a 3' untranslated region of 217 bp, and an open-reading frame of 1,983 bp encoding a polypeptide of 660 amino acids with an estimated molecular weight of 71.39 kDa and an estimated isoelectric point of 5.03. EPHsp70 had five degenerate repeats of tetrapeptide GGMP and three typical Hsp70 signature motifs. The C-terminus amino acid sequence of cytosolic EPHsp70 was EEVD, and the conservation of EPHsp70 of N-terminus was higher than that of C-terminus. The homology between EPHsp70 and the cytosolic Hsp70s of other algae and land plants was more than 70%.

To establish a molecular-marker-assisted system of breeding and genetic study for Laminaria japonica Aresch., amplified fragment length polymorphism (AFLP) was used to construct a genetic linkage map of L. japonica featuring 230 progeny of F(2) cross population. Eighteen primer combinations produced 370 polymorphic loci and 215 polymorphic loci segregated in a 3:1 Mendelian segregation ratio (P

Ezetimibe inhibits cholesterol absorption in the intestine. Sphingomyelin has strong interactions with cholesterol. We investigated the effects of ezetimibe on Sphingomyelinase (SMase) expression in intestine and liver. After feeding rats with ezetimibe (5 mg/kg per day) for 14 days, acid SMase activities in the liver and in the proximal part of small intestine were reduced by 34 and 25%, respectively. Alkaline SMase (alk-SMase) was increased in the proximal part of the small intestine. Administration of lower doses of ezetimibe reduced acid SMase only in the liver by 14% (P0.05). In cell culture studies, ezetimibe decreased acid SMase activity in Hep G2 and Caco-2 cells dose-dependently. The reductions were more rapid for Hep G2 cells than for Caco-2 cells. Western blot showed that acid SMase protein was decreased in both Hep G2 and Caco-2 cells by 100 microM ezetimibe. The SM content was increased in Hep G2 cells but not Caco-2 cells, and total cholesterol content was increased in both cell lines 24 h after stimulation with 100 microM ezetimibe. Mevastatin, the inhibitor of cholesterol synthesis, induced a mild increase in acid SMase activity in Hep G2 cells but not Caco-2 cells. Following the reduction of acid SMase, ezetimibe at high dose slightly increased alk-SMase activity. In conclusion, the study demonstrates an inhibitory effect of ezetimibe on acid SMase activity and expression in both liver and intestine.

In Laminaria japonica Aresch breeding practice, two quantitative traits, frond length (FL) and frond width (FW), are the most important phenotypic selection index. In order to increase the breeding efficiency by integrating phenotypic selection and marker-assisted selection, the first set of QTL controlling the two traits were determined in F(2) family using amplified fragment length polymorphism (AFLP) and simple sequence repeat (SSR) markers. Two prominent L. japonicas inbred lines, one with "broad and thin blade" characteristics and another with "long and narrow blade" characteristics, were applied in the hybridization to yield the F(2) mapping population with 92 individuals. A total of 287 AFLP markers and 11 SSR markers were used to construct a L. japonica genetic map. The yielded map was consisted of 28 linkage groups (LG) named LG1 to LG28, spanning 1,811.1 cM with an average interval of 6.7 cM and covering the 82.8% of the estimated genome 2,186.7 cM. While three genome-wide significant QTL were detected on LG1 (two QTL) and LG4 for "FL," explaining in total 42.36% of the phenotypic variance, two QTL were identified on LG3 and LG5 for the trait "FW," accounting for the total of 36.39% of the phenotypic variance. The gene action of these QTL was additive and partially dominant. The yielded linkage map and the detected QTL can provide a tool for further genetic analysis of two traits and be potential for maker-assisted selection in L. japonica breeding.

In this study, a full-length cytosolic heat shock protein 70 complementary DNA (cDNA) of Laminaria japonica (designated as LJHsp70) was obtained by reverse transcriptase-polymerase chain reaction (RT-PCR) coupled with rapid amplification of cDNA ends. The full length of LJHsp70 cDNA was 2,918 bp, with a 5' untranslated region of 248 bp, a 3' untranslated region of 696 bp, and an open reading frame of 1,974 bp encoding a polypeptide of 657 amino acids with an estimated molecular mass of 72.03 kDa and an estimated isoelectric point of 4.97. There was highly repeated sequence of CAA in 5' untranslated region of LJHsp70. The result of phylogenetic tree of Hsp70s, the BLAST program, analysis and cytosolic Hsp70-specific motif of LJHsp70 verified that the cloned LJHsp70 belonged to cytosolic Hsp70 family. Three typical Hsp70 signature motifs were detected in LJHsp70 by InterPro analysis. Under different stress conditions, messenger RNA (mRNA) expression levels of LJHsp70 were quantified by quantitative RT-PCR. To L. japonica sporophytes kept in different temperatures for 1 h, the expression level of LJHsp70 at 30 degrees C was highest and twofold higher than that at 10 degrees C. To L. japonica sporophytes kept at 25 degrees C for different times, the mRNA expression level of LJHsp70 reached a maximum level after 7 h and then dropped progressively. The expression level of LJHsp70 at 0 or 5 per thousand salt concentration for 2 h was twofold higher than that at 30 per thousand salt concentration for 2 h. The results showed that LJHsp70 may be a kind of potential biomarker used to monitor environment conditions.

Effect of temperature and irradiance on growth and reproduction of Enteromorpha prolifera that bloomed offshore along the Qingdao coast in summer 2008, was studied. It was showed that E. prolifera propagated mainly asexually with specific growth rate (SGR) of 10.47 at 25°C/40 μmol m−2s−1. Under this condition, gametes with two flagellate formed and released in 5 days. At the beginning of the development, the unicell gamete divided into two cells with heteropolarity, and then the apical cell developed into thalli primordial cells, whereas the basal cell developed into rhizoid primordial cells. In 8-day culture, the monoplast gamete developed into juvenile germling of 240 μm in length. Unreleased gametes can develop directly within the alga body. E. prolifera could either reproduce through lateral branching or fragmenting except apomixis revealed by Microscopic observation. On aged tissue of E. prolifera, although the degraded pigments partially remained in faded algal filaments, numerous vegetative cells could still divide actively in the algal tissues.